D. vulgaris Cascade/I-C (Cas5c-Cas8c-Cas7)/pHMGWA
(Plasmid #81185)

• Purpose: Expresses Desulfovibrio vulgaris Cascade/I-C complex subunits in E. coli with TEV-cleavable N-terminal His-MBP tag on Cas5c
• Depositing Lab: Jennifer Doudna
• Publication: Hochstrasser et al Mol Cell. 2016 Sep 1;63(5):840-51. doi: 10.1016/j.molcel.2016.07.027.

Backbone

• Vector backbone: pHMGWA
• Backbone manufacturer: ThermoFisher
• Backbone size w/o insert (bp): 8236
• Total vector size (bp): 10041
• Vector type: Bacterial Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: Cas5c
• Alt name: Cas5d
• Alt name: Cas5
• Species: Desulfovibrio vulgaris str. Hildenborough
• Insert Size (bp): 680
• GenBank ID: 2781521 YP_009170
• Promoter: T7
• Tag / Fusion Protein:
  • 6xHis - MBP - TEV protease site (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gateway Cloning
• 5′ sequencing primer: MBP forward
• 3′ sequencing primer: T7 terminator

Gene/Insert 2

• Gene/Insert name: Cas8c
• Alt name: Csd1
• Species: Desulfovibrio vulgaris str. Hildenborough
• Insert Size (bp): 1839
• GenBank ID: 2781582 YP_009171.1
• Promoter: T7

Cloning Information for Gene/Insert 2

• Cloning method: Gateway Cloning
• 5′ sequencing primer: MBP forward
• 3′ sequencing primer: T7 terminator

Gene/Insert 3

• Gene/Insert name: Cas7
• Alt name: Csd2
• Species: Desulfovibrio vulgaris str. Hildenborough
• Insert Size (bp): 873
• GenBank ID: 2781595 YP_009172.1
• Promoter: T7

Cloning Information for Gene/Insert 3

• Cloning method: Gateway Cloning
• 5′ sequencing primer: MBP forward
• 3′ sequencing primer: T7 terminator

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Entire set of cas5c-cas8c-cas7 genes amplified together from genomic D. vulgaris DNA and TOPO/Gateway cloned into pHMGWA destination vector. A single T7 promoter drives expression of all three genes.